Epstein-BARR virus nucleic acid constructs and vaccines made therefrom, and methods of using same

Abstract

Nucleic acid molecules and compositions comprising one or more nucleotide sequences that encode a consensus Epstein-Barr virus (EBV) antigen. Immunomodulatory methods and methods of inducing an immune response against EBV are disclosed. Method of treating infection by EBV and methods of treating or preventing a disease or disorder associated with EBV are disclosed. Modified consensus EBV antigens are disclosed.

What is claimed is: 
     
         1 . An immunogenic composition comprising a nucleic acid molecule encoding at least one Epstein-Barr virus (EBV) antigen,
 wherein the nucleic acid molecule encodes at least one peptide comprising an amino acid sequence selected from the group consisting of:   a) an amino acid sequence having at least 90% identity over an entire length of the amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO: 26,   b) an immunogenic fragment comprising at least 90% identity over at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO:26,   c) an amino acid sequence elected from the group consisting of SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30 and SEQ ID NO:32,   d) an immunogenic fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO:26.   
     
     
         2 . The immunogenic composition of  claim 1 , wherein the immunogenic composition comprises at least two EBV antigens selected from the group consisting of an EBV glycoprotein antigen, and a latent-stage EBV antigen. 
     
     
         3 . The immunogenic composition of  claim 1 , wherein the immunogenic composition comprises:
 a) at least one EBV glycoprotein antigen;   b) at least one latent-stage EBV antigen; or   c) a combination thereof.   
     
     
         4 . The immunogenic composition of  claim 1 , wherein the nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of
 a) a nucleotide sequence having at least 90% identity over an entire length of a nucleotide sequence selected from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, and SEQ ID NO:25,   b) an immunogenic fragment of a nucleotide sequence having at least 90% identity over at least 60% of nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, and SEQ ID NO:25,   c) a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, and SEQ ID NO:31, and   d) an immunogenic fragment of a nucleotide sequence selected from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, and SEQ ID NO:25.   
     
     
         5 . The immunogenic composition of  claim 1 , wherein
 a) the nucleic acid molecule is selected from the group consisting of a DNA molecule and an RNA molecule encoded by said DNA molecule;   b) the nucleic acid molecule comprises a nucleotide sequence encoding a peptide that is operably linked to at least one regulatory sequence selected from the group consisting of a start codon, a leader sequence, an IgE leader sequence, and at least one stop codon;   c) the nucleic acid molecule comprises an expression vector; and   d) the nucleic acid molecule is incorporated into a viral particle;   or any combination thereof.   
     
     
         6 . The immunogenic composition of  claim 1 , further comprising a pharmaceutically acceptable excipient, an adjuvant, or a combination thereof. 
     
     
         7 . A nucleic acid molecule encoding a peptide comprising an amino acid sequence selected from the group consisting of:
 a) an amino acid sequence having at least 90% identity over an entire length of the amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO:26,   b) an immunogenic fragment comprising at least 90% identity over at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO:26,   c) an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, and SEQ ID NO:32, and   d) an immunogenic fragment comprising at least 60% of an amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO:26.   
     
     
         8 . The nucleic acid molecule of  claim 7 , wherein
 a) the nucleic acid molecule is selected from the group consisting of a DNA molecule and an RNA molecule encoded by said DNA molecule;   b) the encoded peptide is operably linked to at least one regulatory sequence selected from the group consisting of a start codon, a leader sequence, an IgE leader sequence and a stop codon;   c) the nucleic acid molecule comprises an expression vector; and   d) the nucleic acid comprises a viral particle;   or a combination thereof.   
     
     
         9 . The nucleic acid molecule of  claim 7 , wherein the nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of:
 a) a nucleotide sequence having at least 85% identity over an entire length of a nucleotide sequence selected from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, and SEQ ID NO:25,   b) an immunogenic fragment of a nucleotide sequence having at least 85% identity over at least 60% of the nucleotide sequence selected from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, and SEQ ID NO:25,   c) a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, and SEQ ID NO:31,   d) an immunogenic fragment of a nucleotide sequence selected from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, and SEQ ID NO:25.   
     
     
         10 . The peptide of  claim 7 , wherein the peptide is operably linked to an amino acid sequence as set forth in SEQ ID NO:33. 
     
     
         11 . The nucleic acid molecule of  claim 7 , wherein the nucleic acid molecule is operably linked to a nucleotide sequence encoding SEQ ID NO:33. 
     
     
         12 . A peptide, wherein the peptide comprises an amino acid sequence selected from the group consisting of:
 a) an amino acid sequence having at least 90% identity over an entire length of the amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO:26,   b) an immunogenic fragment comprising at least 90% identity over at least 60% of the amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO:26,   c) an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, and SEQ ID NO:32, and   d) an immunogenic fragment comprising at least 60% of an amino acid sequence selected from the group consisting of SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, and SEQ ID NO:26.   
     
     
         13 . A method of inducing an immune response against an EBV antigen or treating a disease or disorder associated with EBV latency in a subject in need thereof, the method comprising administering an immunogenic composition of  claim 1  to the subject. 
     
     
         14 . The method of  claim 13 , wherein the immunogenic composition comprises
 a) at least one EBV glycoprotein antigen;   b) at least one latent-stage EBV antigen; or   c) a combination thereof.   
     
     
         15 . The method of  claim 13 , wherein the disease or disorder associated with EBV latency is at least one of Burkitt's lymphoma, Hodgkin lymphoma, Post-transplant lymphoma, T-cell lymphoma, AIDS-associated B-cell lymphoma, gastric carcinoma, nasopharyngeal carcinoma, infectious mononucleosis, and an autoimmune disease or disorder. 
     
     
         16 . The immunogenic composition of  claim 1  wherein the nucleic acid molecule encodes at least one peptide comprising an amino acid sequence selected from the group consisting of:
 an amino acid sequence having at least 94% identity over an entire length of the amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24,and SEQ ID NO:26.